Abstract
Many Gram-negative pathogens utilize a type III secretion (T3S) system to directly deliver effector molecules into host eukaryotic cells to manipulate cellular processes. These surface-exposed syringe-like structures are highly conserved, necessary for pathogenesis, and hence are therapeutic targets against a number of Gram-negative pathogens. Here we describe a protocol for using purified needle proteins to immunize mice, and subsequently, ways to characterize the immune response to immunization.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Galán JE, Wolf-Watz H (2006) Protein delivery into eukaryotic host cells by type III secretion machines. Nature 444:567–573
Cornelis GR (2006) The type III secretion injectisome. Nat Rev Microbiol 4:811–825
Raymond B, Young JC, Pallett M, Endres RG, Clements A, Frankel G (2013) Subversion of trafficking, apoptosis, and innate immunity by type III secretion system effectors. Trends Microbiol 8:430–4411
Blocker A, Gounon P, Larquet E, Niebuhr K, Cabiaux V, Parsot C, Sansonetti P (1999) The tripartite type III secretion of Shigella flexneri inserts IpaB and IpaC into host membranes. J Cell Biol 147:683–693
Frank DW, Vallis A, Wiener-Kronish JP, Roy-Burman A, Spack EG, Mullaney BP, Megdoud M, Marks JD, Fritz R, Sawa T (2002) Generation and characterization of a protective monoclonal antibody to Pseudomonas aeruginosa PcrV. J Infect Dis 186:64–73
Matson JS, Durick KA, Bradley DS, Nilles ML (2005) Immunization of mice with YscF provides protection from Yersinia pestis infections. BMC Microbiol 5:38
Hill J, Cpose C, Leary S, Stagg AJ, Williamson ED, Titball RW (2003) Synergistic protection of mice against plague with monoclonal antibodies specific for the F1 and V antigens of Yersinia pestis. Infect Immun 71:2234–2238
Health DG, Anderson GW, Mauro JM, Welkos SL, Andrews GP, Adamovicz J, Friedlander AM (1998) Protection against experimental bubonic and pneumonic plague by a recombinant capsular F1-V antigen fusion protein vaccine. Vaccine 16:1131–1137
Leary SEC, Williamson ED, Griffin KF, Russell P, Eley SM, Titball RW (1995) Active immunization with recombinant V antigen from Yersinia pestis protects mice against plague. Infect Immun 63:2854–2858
Martinez-Becerra FJ, Kissmann JM, Diaz-McNair J, Choudhari P, Quick AM, Mellado-Sanchez G, Clements JD, Pasetti MF, Picking WL (2012) Broadly protective Shigella vaccine based on type III secretion apparatus proteins. Infect Immun 80:1222–1231
Jessen DL, Osei-Owusu P, Toosky M, Roughead W, Bradley DS, Nilles ML (2014) Type III secretion needle proteins induce cell signaling and cytokine secretion via toll-like receptors. Infect Immun 82:2300–2309
Osei-Owusu P, Jessen-Condry DL, Toosky M, Roughead W, Bradley DS, Nilles ML (2015) The n terminus of type III secretion needle protein YscF from Yersinia pestis functions to modulate innate immune responses. Infect Immun 83:1507–1522
Moss J, Ehrmantraut ME, Banwart BD, Frank DW, Barbieri JT (2001) Sera from adult patients with cystic fibrosis contain antibodies to pseudomonas aeruginosa type III apparatus. Infect Immun 69:1185–1188
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media New York
About this protocol
Cite this protocol
Alvine, T.D., Bradley, D.S., Nilles, M.L. (2017). Mouse Immunization with Purified Needle Proteins from Type III Secretion Systems and the Characterization of the Immune Response to These Proteins. In: Nilles, M., Condry, D. (eds) Type 3 Secretion Systems. Methods in Molecular Biology, vol 1531. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6649-3_17
Download citation
DOI: https://doi.org/10.1007/978-1-4939-6649-3_17
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6647-9
Online ISBN: 978-1-4939-6649-3
eBook Packages: Springer Protocols